OBJECTIVETo establish a convenient method for diagnosis of lethal avian influenza virus infection.

METHODSNinety-six-well microtiter plates were coated with 100 microl of H5N1 protein (0.1 microg/ml) diluted in coating buffer. These plates were used for enzyme-linked immunosorbent assay (ELISA).

RESULTSFour of the 23 suspected borderline cases of lethal avian influenza virus infection were positive by the ELISA, and all the healthy controls (n=234) were negative. These results were confirmed by hemagglutination-inhibition (HI) and neutralization tests, and results were totally consistent (100 percent) with each other among the three methods.

CONCLUSIONThe ELISA may be used in the screening for lethal avian influenza virus infection.

Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; isolation & purification ; Influenza, Human ; diagnosis ; immunology

OBJECTIVETo explore the effect of cytokine in the different prognosis of patients with severe influenza A (H1N1) infection.

METHODS28 cases with severe influenza A (H1N1) were enrolled in the study including 16 cured cases and 12 dead cases. The cytokine level in serum was detected by Luminex technology.

RESULTSThe levels of IL-2, IL-12 (P70) and IFN-gamma in dead group was lower than cured and normal control group and the difference were significant, P <0.05, respectively. IL-4 level in the dead group was significantly lower than cured group and normal control group, P value was 0.0310 and 0.0012, respectively.

CONCLUSIONSThe Thl cytokine level in the severe 2009 epidemic H1N1 influeaze cases shows decreased trend, and the trend is more obvious in dead cases. The decrease of Th1 cytokine may be one of reasons leading to severe clinical situation and related withthe bad prognosis.

Cytokines ; blood ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; isolation & purification ; Influenza, Human ; diagnosis ; immunology ; mortality ; virology ; Prognosis ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo understand the infection condition and analytical methods of Influenza A (H1N1) virus in the population of Hunan Province during different periods.

METHODSQuick surveys on the positive rate of Influenza A (H1N1) virus hemagglutination inhibition (HI) test have been conducted for 5 times successively from November 2009 to March 2010 in 14 medical and health institutions of Changsha city, whose results were then compared with those from the sampling surveys of whole Hunan province.

RESULTS2131 subjects were involved in this study; the total population standardized rates of antibody positive investigated for 5 times were 9.32% , 14.62%, 31.08%, 28.43% and 22.80% respectively; the population of 6-17-years-old has the highest rate of antibody positive; only 9.84% of the antibody positive subjects attributed to vaccine inoculation; there was no significant difference in the standardized positive rates between the quick serological surveys and the corresponding sampling survey of Hunan province (P > 0.05).

CONCLUSIONThe positive rate of A (H1N1) virus antibody reached the peak in late January 2010; quick investigations in small region could be used to evaluate the infection prevalence during pandemic of infectious diseases.

Adolescent ; Adult ; Antibodies, Viral ; blood ; Child ; China ; Female ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; diagnosis ; Male ; Middle Aged ; Vaccination

BACKGROUNDTo understand the optimal condition of single radial hemolysis (SRH) for diagnosis of avian influenza A (H5N1) virus in order that SRH could be performed in general laboratories.

METHODSThe effect of different concentration of virus and species of red blood cells, as well as kind and concentration of agarose on testing sensitivity of SRH was determined. Meanwhile the sensitivity and specificity of this method were compared with those of micro-neutralization test.

RESULTSThe optimal condition of SRH included the viral concentration of 1000 HA units per 0.1 ml packed chicken red blood cells, the agarose concentration of 1.0%, the compliment added into agarose-virus-rbc slides after diffusion of sera. The sensitivity and specificity of SRH were very similar to those of micro-neutralization test. Meanwhile, no cross reaction between antibodies, especially antibodies against N1 antigens, H5N1 and H1N1 viruses was detected.

CONCLUSIONThe sensitivity and specificity of SRH were very similar to those of micro-neutralization assay. SRH could be performed in normal laboratories and be used for testing large scale serum samples.

Animals ; Antibodies, Viral ; analysis ; Chick Embryo ; Guinea Pigs ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza, Human ; diagnosis ; immunology ; Neutralization Tests ; Orthomyxoviridae Infections ; diagnosis ; immunology

OBJECTIVETo investigate the epidemiological features of influenza virus B (IVB) in the winter and the clinical features of pediatric pneumonia caused by IVB only.

METHODSA retrospective study was performed on the clinical data of children with respiratory infection who received pathogen testing and therapy at Soochow University Affiliated Children's Hospital during the winters of 2008, 2009, 2010 and 2011.

RESULTSThe positive rates of influenza viruses A and B in the winters of 2008, 2009, and 2010 were 0.89%, 5.49%, and 6.24% respectively; the positive rate of influenza viruses A and B in the winter of 2011 was 8.72%, significantly higher than those in 2008-2010. The positive rates of IVB in the winters of 2008, 2009, and 2010 were 0%, 0%, and 0.21% respectively; the positive rate of IVB in the winter of 2011 was 5.36%, which was significantly higher than in the years 2008 to 2010. Pneumonia caused by IVB was confirmed in 94 children during the winter of 2011, including 27 cases of pneumonia caused by IVB only. Most of children with pneumonia caused by IVB only were aged over 6 months. The common symptoms in the 27 children caused by IVB only were fever (85%), runny nose (89%), and cough (100%). Wheezing (26%) and dyspnea (7%) were also seen in some cases. Among the 27 children, 19% showed abnormal white blood cell count, 30% showed increased C-reactive protein, 70% showed decreased prealbumin, and none showed visible organ dysfunction. No specific imaging findings were seen in the children with pneumonia caused by IVB only. However, many abnormal humoral and cellular immunological parameters were found in the majority of these children. The average length of hospital stay was approximately one week, there were no critical patients and the prognosis was good.

CONCLUSIONSInfluenza viruses were at a peak level in inpatient children in the winter of 2011. IVB infection rate was gradually increasing. In children with pneumonia caused by IVB only, there are few critical patients, the symptoms are nonspecific and the prognosis is good.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Influenza B virus ; Influenza, Human ; diagnosis ; epidemiology ; immunology ; Length of Stay ; Male ; Pneumonia, Viral ; diagnosis ; epidemiology ; immunology ; Retrospective Studies

Though the 2009 worldwide influenza A (H1N1) pandemic has been declared to have ended, the influenza virus is expected to continue to circulate from some years as a seasonal influenza. A rapid antigen test (RAT) can aid in rapid diagnosis and allow for early antiviral treatment. We evaluated the clinical usefulness of RAT using SD Bioline Influenza Antigen Test(R) kit to detect the influenza virus, considering various factors. From August 1, 2009 to October 10, 2009, a total of 938 patients who visited the outpatient clinic at Korea University Guro Hospital with influenza-like illnesses were enrolled in the study. Throat or nasopharyngeal swab specimens were obtained from each of the patients. Using these specimens, we evaluated the influenza detection rate by rapid antigen test based on the real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) method. In comparison with rRT-PCR, the sensitivity and specificity of the RAT were 44.0% and 99.9%, respectively. The cyclic threshold values of RAT negative specimens were higher than RAT positive specimens (30.1+/-3.1 vs. 28.3+/-3.9, p=0.031). The sensitivity of the RAT kit was higher in patients who visited clinics within two days of symptom onset (60.4% vs. 11.1%, p=0.026). The results of this study show that the RAT cannot be recommended for general use in all patients with influenza-like illness because of its low sensitivity. The RAT may be used, only in the settings with limited diagnostic resources, for patients who visit a clinic within two days of symptom onset.
Antigens, Viral/genetics ; Humans ; Influenza A Virus, H1N1 Subtype/genetics/immunology/*isolation & purification ; Influenza, Human/*diagnosis/virology ; Reagent Kits, Diagnostic ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Time Factors

OBJECTIVETo study the clinical characteristics and laboratory results of 24 confirmed H1N1 influenza cases.

METHODSThe characters of clinical, laboratory, iconography and etiology of 24 patients with A/H1N1 were studied, and the changes of T-lymphocyte subsets that between the pre- and post-treatment were evaluated.

RESULTSThe ages of patients were ranged from 6 to 65 years old; average age was 26 years old.15 patients were under 25 years old. 22 (22/24, 91.7%) patients had recently traveled to USA or Canada. The most common presenting symptoms were: fever (22/24, 91.7%); sore throat (22/24, 91.7%); cough (20/24, 83.3%); dry cough (14/24, 58.3%); expectoration (6/24, 25.0%); nasal discharge (6/24, 25.0%). Six had pneumonia in sixteen patients (6/16, 37.5%) who took CT scan; seven (7/24, 29.2%) had headache and four (4/24, 16.7%) had muscular soreness; two (2/24, 8.3%) had sneeze and nasal obstruction; only one(1/24, 4.2%) had diarrhea; one (1/24, 4.2%) had conjunctivitis. The result of 23 patients about T-Lymphocyte subsets: most of CD4 and CD8 were decreased (18/23, 78.3%), ranging from 122 to 691 cells/microl (normal was 706 - 1125 cells/microl), with the average of 408 cells/microl, but ratios of CD4/CD8 were normal. Fourteen patients were detected CD4 and CD8 after received the treatment during 5 to 7 days.the results of CD4 (cells/microl) were different between the pre- and post-treatment: 436.29 +/- 189.06, 976.71 +/- 332.96 (paired-samples t test: t = -5.416, P < 0.05) while the results of CD8 (cells/microl) were: 323.64 +/- 176.47, 703.14 +/- 211.77 (t = -5.319, P < 0.05); the results of leukocytes in 22 patients were different between pre- and post-treatment: (5.13 +/- 1.47) x 10(9)/L, (6.25 +/- 1.37) x 10(9)/L (t = -2.900, P < 0.05) while the results of lymphocytes were: (1.16 +/- 0.43) x 10(9)/L, (2.30 +/- 0.37) x 10(9)/L (t = -6.819, P < 0.05); but the ratios of CD4/CD8 were: 1.44 +/- 0.41, 1.40 +/- 0.26 (t = 0.507, P > 0.05). All the patients were received antivirus treatment (Oseltamivir) and the virus conversed during 1 - 10 days (average 4.5 days). The temperature was normal after onset during 3 - 4 days and the patients were recovered during 3 - 13 days (with the average of 7.3 days).

CONCLUSIONInfluenza A virus H1N1 subtype was identified as the cause of outbreaks of febrile respiratory infection which was self-limited. There was no evidence to show that the changes of T-Lymphocyte subsets could indicate the prognosis of patients.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; diagnosis ; immunology ; virology ; Male ; Middle Aged ; Prognosis ; T-Lymphocyte Subsets ; Young Adult

Uveitis was a rare adverse event of vaccination. We met two cases of acute uveitis with exudative retinal detachment following vaccination of H1N1 influenza. Case 1 was a 10-year-old boy who was admitted for bilateral blurred vision at 10 days after vaccination of H1N1 influenza. Vitreous opacity was obvious in both eyes. Broad exudative retinal detachment was observed in the right eye. Case 2 was a 47-year-old female who suffered from an acute high fever at 2 days after the vaccination of H1N1 influenza. Later, she encountered bilateral headache and decreasing vision. In both eyes, mutton fat keratic precipitates, positive Tyndall phenomenon, congestion of optic disc and exudative retinal detachment were observed.
Child ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; pathogenicity ; Influenza Vaccines ; adverse effects ; therapeutic use ; Influenza, Human ; immunology ; prevention & control ; Male ; Middle Aged ; Retinal Detachment ; diagnosis ; etiology ; Uveitis ; diagnosis ; etiology

Inflammatory bowel disease (IBD) is a long-standing disease that often requires long-term use of immunosuppressive agents including immunomodulators (such as azathioprine, 6-mercaptopurine and methotrexate) and tumor necrosis factor-alpha inhibitors (such as infliximab and adalimumab). Introduction of immunosuppressive therapies, however, involves the risk of host susceptibility to opportunistic infections in this patient population. Therefore, adequate immunization for vaccine-preventable infectious diseases is currently recommended for all patients with IBD and is emerging as an important target for quality improvements in IBD care. However, ongoing issues regarding underuse of immunization, safety and efficacy of vaccines in patients with IBD remain. For quality improvements in IBD care, all physicians should follow the recent immunization guidelines proposed by professional IBD societies. Additionally, there are ongoing needs for intensive educational programs regarding a role of immunization in long-term care of IBD and up-to-date immunization guidelines. Immunization status should be checked at the time of diagnosis of IBD and timely vaccination before initiation of immunosuppressive therapies can be a practical solution for maximizing the efficacy of vaccination at this point. Inactivated vaccines can be used safely irrespective of immunization status of patients, while attenuated vaccines are contraindicated in patients on immunosuppressive therapies. This article reviews an ideal strategy for vaccinating patients with IBD based on the currently recommended immunization guidelines.
Antibodies, Monoclonal/therapeutic use ; Humans ; Immunosuppressive Agents/therapeutic use ; Inflammatory Bowel Diseases/diagnosis/drug therapy/*immunology ; Influenza Vaccines/immunology ; Influenza, Human/prevention & control ; Pneumonia/prevention & control ; *Vaccination ; Vaccines, Synthetic/immunology

OBJECTIVETo ascertain the causation of a family cluster involving two undefined pneumonia cases, a 12-year-old girl and her brother, reported October, 2005 in Xiangtan county, Hunan province.

METHODSInformation on epidemiology and clinical manifestation of the cases was collected from interviewing the keyman and referring to related medical records. The environment exposure of the cases to their households and the timeline of the illness were reproduced, using this information. Medical check-up was undergone among the close contacts of the cases and on sick/dead poultry. Throat swab of the cases were collected and tested by both RT-PCR and real-time PCR to detect viral nucleic acids of A/H5N1, and were then inoculated into special pathogen free (SPF) embryonated hens' eggs. Serum of the cases including acute and convalescent phases were also collected and tested by microneutralization and haemagglutination-inhibition (HI) assays to detect H5-specific antibodies.

RESULTSBoth the girl and her brother developed fever 2 and 4 days after sudden deaths of chickens being raised in the same house. Both of them had developed pneumonia and the girl died from acute respiratory distress syndrome (ARDS) complicated with multi-organ failure. The boy survived and subsequently discharged from hospital. An eighth-day serum from the girl tested H5 antibody negative, while 4-fold and greater increased in antibody titers were detected in serum from the boy using microneutralization and HI assays in sequential acute and convalescent sera. Of 192 cases, only one doctor who cared for the girl during hospitalization had upper respiratory symptoms but tested negative for H5N1 by microneutralization assay.

CONCLUSIONThe boy was the first confirmed human case of avian influenza A (H5N1) in the mainland of China and his sister was diagnosed clinically. The most probable explanation of these two cases was that the transmission of H5N1 virus from infected poultry within the same household environment. No evidence of human-to-human transmission was noted in the family cluster.

Animals ; Chickens ; Child ; China ; Fatal Outcome ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; isolation & purification ; Influenza in Birds ; transmission ; Influenza, Human ; complications ; diagnosis ; transmission ; Male ; Pneumonia ; virology ; Respiratory Distress Syndrome, Adult ; virology